Abstract Seven Pakistani brown rice cultivars (KS-282, SB, C-9, SUPRI, PK-386, IRRI-9 and IRRI-6) were investigated as an alternative raw material to develop directly expanded extrudates. Results indicated that bulk density remained constant in all rice cultivars, whereas SUPRI presented a higher length-breadth (LB) ratio and lower thousand kernel weight and starch content than other cultivars. Amylose content was higher in KS-282, C-9 and IRRI-9 and lowest in SB. SUPRI rice extrudates produced optimum expansion and crispiness with reduced hardness, piece and bulk density. Scanning electron micrographs showed more cell pockets and evenness in porosity per unit area in SUPRI brown rice extrudates, indicating more porous and crispier extrudates. It is also evidenced from the micrographs that cavities were evenly distributed and less ruptured in SUPRI brown rice. Consequently, SUPRI extrudates exhibited better microstructure, crispiness (No. of peaks 9), and porosity (0.838 ± 0.009) along with minimum piece density (0.038 ± 0.001 g/cm3) and hardness (17.85 ± 1.05 N). Conclusively, low amylose rice cultivar (SUPRI) perform better as substitute raw material for extrudates development.

Abstract Six protein energy-bars (B1-B6) were prepared for Pakistani-athletes using dates, dried apricots, Cheddar-cheese and whey-protein isolate. Bars B1-B3 contained 5 g Cheddar-cheese and 13 g whey-protein isolate while quantity of dates were 74, 68 and 65 g and apricots 8, 14 and 17 g respectively. Bars B4-B6 contained 8g Cheddar-cheese and 12 g whey protein isolate while dates and apricots were same as in bars B1-B3. Bars were sealed in aluminium foil, stored at 20 ± 5 °C for 45 days to evaluate their sensory, compositional, physicochemical, microbial, textural and antioxidant properties. All bars have good sensory attributes except B6 that showed (p < 0.05) lower acceptability. The pH, water activity and microbial count decreased (p < 0.05) in all the bars. Maximum hardness was noticed in bar B1 and lowest in B6 while highest firmness was recorded in B2 and lowest in B6. The increase (p < 0.05) of total phenolic content was noticed in B6 and the highest increase of total flavonoid content was estimated in B2 at day 45, while B5 showed the highest antioxidant-activity on the 30th day afterwards a decline was observed in all the bars. Study revealed that these bars have good sensory attributes, physical characteristics and shelf-stability and could be a healthy snack for athletes.

Proteins and peptides are the most diverse biomolecules found in nature and make our interest due to their wide applications in food and pharmaceutical industry. Angiotensin Converting Enzyme (ACE) plays a major role in controlling blood pressure. The inhibition of ACE with peptides is a main target in the regulation of hypertension. The objective of the present study was to investigate the therapeutic potential of soy bean. This was accomplished by isolation of ACE inhibitory peptides using response surface methodology (RSM) and characterization of these bioactive peptides by mass spectrometry. 31 hydrolyzed fractions were isolated and evaluated for their ACE inhibition potential. Hydrolyzed fraction having highest ACE inhibitory activity was characterized by liquid chromatography-mass spectrometry (LC-MS) technique. RSM results showed maximum ACE inhibition potential (64%) by hydrolyzate was obtained at 45 ºC temperature, pH 8.0, E/S 0.2 in 2 hours hydrolysis time. Results of LC-MS analysis revealed Ser-Gly, Ser-Pro, Met-Ala, His-Ala, Lys-Pro, Phe-Thr, Met-Leu, Pro-Arg, Ala-Pro-Val, Pro-Ala-Leu, Val-Met-Gly, Pro-Leu-Val, Pro-Pro-Gln, His-Arg-Gly, Ser-Phe-Val-Leu, Ala-Val-His-Try, Arg-Thr-Val-Arg, His-His-Tyr-Leu-Val, Asp-Gly-Ala-Cys-Ser-Ala-Asn and MetVal-Thr-Gly-Pro-Gly-Cys-His bioactive peptides in hydrolyzed fraction of soy bean. Our data provide evidence that response surface methodology is a good approach for isolation of antihypertensive bioactive peptides with more potent activity as nutraceuticals or pharmaceuticals. Therefore soy bean can be use for industrial production of pharmaceutical grade natural medicines for handling high blood pressure.

Abstract Streptokinase (SK) is an enzyme that is used for the treatment of cardiovascular diseases. The current study focused on the enhanced production of SK by inducing mutation in Streptococcus agalactiae EBL-20 and optimization of medium components and culture conditions for the maximum growth of mutant derived strain. S. agalactiae EBL-32 was selected as a potent mutant after exposure of S. agalactiae EBL-20 to EMS for 180 minutes. SK activity obtained from mutant derived strain was found to be 1.6 fold higher as compared to the activity achieved by wild strain. Nutritional requirements of the mutated strain were optimized by single factor analysis method suggesting glucose as the optimum carbon source; yeast extract and peptone as a suitable nitrogen sources and corn steep liquor (CSL) as an appropriate substrate for the maximum SK production. The culture conditions determined by response surface methodology (RSM) suggested that a temperature value of 37.5⁰C and pH 7 of the fermentation medium with 2.50 mL inoculum size for 36 hours of incubation was optimum for maximum yield of SK. Hence the optimization studies resulted into 1.92 fold increase in the yield of SK suggesting the new isolate suitable for commercial scale production of SK.

This work aimed to study the production and purification of glucose oxidase by Aspergillus niger and Penicillium notatum using corn steep liquor as the substrate and evaluate its antimicrobial activity for use in pharmaceutical and food industries. The enzyme was purified by ammonium sulfate precipitation (60-85%), DEAE-cellulose ion exchange and Sephadex G-200 size exclusion chromatography. The crude enzyme extracts of A. niger and P. notatum showed 2.32 and 5.53 U mg-1 specific activities, respectively, which after desalting was 15.52 and 12.05 U mg-1, and after ion exchange and gel filtration chromatography was 29.09 - 62 and 25.72 - 59.37 U mg-1 for A. niger and P. notatum, respectively. The antimicrobial activity was determined by disc diffusion method against selected microbial strains where glucose oxidase from A. niger showed anti-bacterial activity, while no fungicidal effects were shown by both A. niger and P. notatum glucose oxidases.

In the present study enhanced production of glucose aerodehydrogenase from Aspergillus niger has been achieved after optimizing the dose of chemical mutagen ethyl methane sulfonate (EMS) that has not been reported earlier. Different doses of mutagen were applied and a strain was developed basing upon the best production. The selected strain Aspergillus niger EMS-150-F was optimized for nutrient requirements in order to produce enzyme through fermentation and the results showed the best yield at 2% corn steep liquor (CSL), 36 hours fermentation time, pH 5, 30°C temperature, 0.3% KH2PO4, 0.3% urea and 0.06% CaCO3. The enzyme was then purified and resulted in 57.88 fold purification with 52.12% recovery. On kinetic characterization, the enzyme showed optimum activity at pH 6 and temperature 30°C. The Michaelis-Menton constants (Km, Vmax, Kcat and Kcat/Km) were 20 mM, 45.87 U mL-1, 1118.81 s-1 and 55.94 s-1 mM-1, respectively. The enzyme was found to be thermaly stable and the enthalpy and free energy showed an increase with increase in temperature and ΔS* was highly negative proving the enzyme from A. niger EMS-150-F resistant to temperature and showing a very little disorderliness.

Glucose oxidase is an active oxygen species generating enzyme produced from Aspergillus niger grown in submerged fermentation. Disintegration of the mycelium resulted in high glucose oxidase activity that was subjected to ammonium sulfate precipitation at 60-85% saturation rates that resulted to 6.14 U mg -1 specific activity. Purification of enzyme by anion exchange column (DEAE-Cellulose) resulted into 22.53 U mg-1 specific activity and 10.27 fold purification. This was applied to sephadex G-200 column for gel filtration chromatography. It was observed that enzyme achieved 59.37 U mg-1of specific activity with 27.08 fold purity and 64.36% recovery. Purified glucose oxidase was injected into rabbits through intravenous route, to raise the glucose oxidase antibodies. After 30 days incubation period, the rabbits were slaughtered and serum was separated from blood. The antibodies were isolated by ammonium sulfate precipitation and confirmed by agar gel precipitation test. This could be a convenient and low cost alternate assay for the estimation of glucose oxidase in biological fluids. Moreover, such antibodies against the said enzyme could be used in various therapeutic and diagnostic applications.

The present investigation deals with the kinetics of submerged extracellular lipases fermentation by both wild and mutant strains of Rhizopus oligosporus var. microsporus in a laboratory scale stirred fermentor. Other parameters studied were inoculum size, pH, agitation and rate of aeration. It was found that the growth and lipases production was increased gradually and reached its maximum 9.07± 0.42ª U mL-1 (W) and 42.49 ± 3.91ª U mL-1 (M) after 30h of fermentation for both wild and mutant strain. There is overall increase of 109% (W) and 124% (M) in the production of extracellular lipases as compared to shake flask. Another significant finding of the present study is that the fermentation period is reduced to 30 h in case of wild and 23 h in case of mutant from 48 h in shake flask studies. The specific productivity of mutant strain (qp = 377.3 U/g cells/h) was several folds higher than wild strain. The specific production rate and growth coefficient revealed the hyperproducibility of extracellular lipases using mutant IIB-63NTG-7.

Comparative efficacy of turmeric (Curcuma longa L.) crude powder and salinomycin sodium on the occurrence of coccidiosis and growth performance of broiler was evaluated. A total of 90, day-old chicks were randomly divided into six groups. From first day onward, ration was supplemented with 1, 2 and 3 % turmeric powder in groups 2, 3 and 4, respectively, group 1 received salinomycin sodium @ 12 g 50 kg-1 feed while groups 5 and 6 were kept as infected un-medicated and uninfected un-medicated controls. First five groups were infected with Eimeria tenella sporulated oocysts @ 1,00,000/chick at the age of 20 days. Body weight gain, feed consumption, feed conversion ratio were investigated throughout the experimental period, and bloody diarrhea and oocysts excretions were investigated at the first and the second week after infection. Maximum coccidiostatic effect was observed with turmeric (3%) showing mild bloody diarrhea as compared to other infected groups receiving turmeric containing rations. This effect was comparable with a standard coccidiostat i.e., salinomycin sodium. Similarly, the weight gain in the groups treated with salinomycin sodium (2280g) and 3% turmeric (2293g) were also significantly higher (p < 0.05) than that of infected control group (1955g). In the groups treated with ration supplemented with 3% turmeric powder and salinomycin sodium, the peak excretion of oocysts was delayed about 1 or 2 days relative to the control infected group. Concentration-dependent coccidiostatic effect of turmeric suggested that further studies should be carried out to determine the possible maximum safe levels of turmeric with least toxic effects to be used as coccidiostat.